| First Author | Pyrshev K | Year | 2022 |
| Journal | FASEB J | Volume | 36 |
| Issue | 5 | Pages | e22275 |
| PubMed ID | 35349181 | Mgi Jnum | J:349393 |
| Mgi Id | MGI:7355067 | Doi | 10.1096/fj.202200160R |
| Citation | Pyrshev K, et al. (2022) ClC-K2 Cl(-) channel allows identification of A- and B-type of intercalated cells in split-opened collecting ducts. FASEB J 36(5):e22275 |
| abstractText | The collecting duct is a highly adaptive terminal part of the nephron, which is essential for maintaining systemic homeostasis. Principal and intercalated cells perform different physiological tasks and exhibit distinctive morphology. However, acid-secreting A- and base secreting B-type of intercalated cells cannot be easily separated in functional studies. We used BCECF-sensitive intracellular pH (pHi ) measurements in split-opened collecting ducts followed by immunofluorescent microscopy in WT and intercalated cell-specific ClC-K2(-/-) mice to demonstrate that ClC-K2 inhibition enables to distinguish signals from A- and B-intercalated cells. We show that ClC-K2 Cl(-) channel is expressed on the basolateral side of intercalated cells, where it governs Cl(-) -dependent H(+) /HCO3 (-) transport. ClC-K2 blocker, NPPB, caused acidification or alkalization in different subpopulations of intercalated cells in WT but not ClC-K2(-/-) mice. Immunofluorescent assessment of the same collecting ducts revealed that NPPB increased pHi in AE1-positive A-type and decreased pHi in pendrin-positive B-type of intercalated cells. Induction of metabolic acidosis led to a significantly augmented abundance and H(+) secretion in A-type and decreased proton transport in B-type of intercalated cells, whereas metabolic alkalosis caused the opposite changes in intercalated cell function, but did not substantially change their relative abundance. Overall, we show that inhibition of ClC-K2 can be employed to discriminate between A- and B-type of intercalated cells in split-opened collecting duct preparations. We further demonstrate that this method can be used to independently monitor changes in the functional status and abundance of A- and B-type in response to systemic acid/base stimuli. |
